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. 1997 Feb;179(4):1035–1043. doi: 10.1128/jb.179.4.1035-1043.1997

Altered transcription activation specificity of a mutant form of Bacillus subtilis GltR, a LysR family member.

B R Belitsky 1, A L Sonenshein 1
PMCID: PMC178795  PMID: 9023181

Abstract

A mutation (gltR24) that allows Bacillus subtilis glutamate synthase (gltAB) gene expression in the absence of its positive regulator, GltC, was identified. Cloning and sequencing of the gltR gene revealed that the putative gltR product belongs to the LysR family of transcriptional regulators and is thus related to GltC. A null mutation in gltR had no effect on gltAB expression under any environmental condition tested, suggesting that gltR24 is a gain-of-function mutation. GltR24-dependent transcription of gltAB, initiated at the same base pair as GltC-dependent transcription, was responsive to the nitrogen source in the medium and required the integrity of sequences upstream of the gltAB promoter that are also necessary for GltC-dependent expression. Expression of the gltC gene, transcribed divergently from gltA from an overlapping promoter, was not affected by GltR. Both wild-type GltR and GltR24 negatively regulated their own expression. The gltR gene was mapped to 233 degrees on the B. subtilis chromosome, very close to the azlB locus.

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Selected References

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  1. Ahmed M., Lyass L., Markham P. N., Taylor S. S., Vázquez-Laslop N., Neyfakh A. A. Two highly similar multidrug transporters of Bacillus subtilis whose expression is differentially regulated. J Bacteriol. 1995 Jul;177(14):3904–3910. doi: 10.1128/jb.177.14.3904-3910.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Azevedo V., Alvarez E., Zumstein E., Damiani G., Sgaramella V., Ehrlich S. D., Serror P. An ordered collection of Bacillus subtilis DNA segments cloned in yeast artificial chromosomes. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6047–6051. doi: 10.1073/pnas.90.13.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Belitsky B. R., Janssen P. J., Sonenshein A. L. Sites required for GltC-dependent regulation of Bacillus subtilis glutamate synthase expression. J Bacteriol. 1995 Oct;177(19):5686–5695. doi: 10.1128/jb.177.19.5686-5695.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Belitsky B. R., Sonenshein A. L. Mutations in GltC that increase Bacillus subtilis gltA expression. J Bacteriol. 1995 Oct;177(19):5696–5700. doi: 10.1128/jb.177.19.5696-5700.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bohannon D. E., Rosenkrantz M. S., Sonenshein A. L. Regulation of Bacillus subtilis glutamate synthase genes by the nitrogen source. J Bacteriol. 1985 Sep;163(3):957–964. doi: 10.1128/jb.163.3.957-964.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bohannon D. E., Sonenshein A. L. Positive regulation of glutamate biosynthesis in Bacillus subtilis. J Bacteriol. 1989 Sep;171(9):4718–4727. doi: 10.1128/jb.171.9.4718-4727.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dubnau D., Davidoff-Abelson R. Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol. 1971 Mar 14;56(2):209–221. doi: 10.1016/0022-2836(71)90460-8. [DOI] [PubMed] [Google Scholar]
  11. Everett M., Walsh T., Guay G., Bennett P. GcvA, a LysR-type transcriptional regulator protein, activates expression of the cloned Citrobacter freundii ampC beta-lactamase gene in Escherichia coli: cross-talk between DNA-binding proteins. Microbiology. 1995 Feb;141(Pt 2):419–430. doi: 10.1099/13500872-141-2-419. [DOI] [PubMed] [Google Scholar]
  12. Falcone D. L., Tabita F. R. Complementation analysis and regulation of CO2 fixation gene expression in a ribulose 1,5-bisphosphate carboxylase-oxygenase deletion strain of Rhodospirillum rubrum. J Bacteriol. 1993 Aug;175(16):5066–5077. doi: 10.1128/jb.175.16.5066-5077.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fouet A., Sonenshein A. L. A target for carbon source-dependent negative regulation of the citB promoter of Bacillus subtilis. J Bacteriol. 1990 Feb;172(2):835–844. doi: 10.1128/jb.172.2.835-844.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glaser P., Kunst F., Arnaud M., Coudart M. P., Gonzales W., Hullo M. F., Ionescu M., Lubochinsky B., Marcelino L., Moszer I. Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325 degrees to 333 degrees. Mol Microbiol. 1993 Oct;10(2):371–384. [PubMed] [Google Scholar]
  15. Goethals K., Van Montagu M., Holsters M. Conserved motifs in a divergent nod box of Azorhizobium caulinodans ORS571 reveal a common structure in promoters regulated by LysR-type proteins. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1646–1650. doi: 10.1073/pnas.89.5.1646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Guérout-Fleury A. M., Frandsen N., Stragier P. Plasmids for ectopic integration in Bacillus subtilis. Gene. 1996 Nov 21;180(1-2):57–61. doi: 10.1016/s0378-1119(96)00404-0. [DOI] [PubMed] [Google Scholar]
  17. He M., Wilde A., Kaderbhai M. A. A simple single-step procedure for small-scale preparation of Escherichia coli plasmids. Nucleic Acids Res. 1990 Mar 25;18(6):1660–1660. doi: 10.1093/nar/18.6.1660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Henikoff S., Haughn G. W., Calvo J. M., Wallace J. C. A large family of bacterial activator proteins. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6602–6606. doi: 10.1073/pnas.85.18.6602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoshino T., Kose-Terai K., Uratani Y. Isolation of the braZ gene encoding the carrier for a novel branched-chain amino acid transport system in Pseudomonas aeruginosa PAO. J Bacteriol. 1991 Mar;173(6):1855–1861. doi: 10.1128/jb.173.6.1855-1861.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Itaya M., Kondo K., Tanaka T. A neomycin resistance gene cassette selectable in a single copy state in the Bacillus subtilis chromosome. Nucleic Acids Res. 1989 Jun 12;17(11):4410–4410. doi: 10.1093/nar/17.11.4410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jin S., Sonenshein A. L. Transcriptional regulation of Bacillus subtilis citrate synthase genes. J Bacteriol. 1994 Aug;176(15):4680–4690. doi: 10.1128/jb.176.15.4680-4690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  23. Kwak J. H., Choi E. C., Weisblum B. Transcriptional attenuation control of ermK, a macrolide-lincosamide-streptogramin B resistance determinant from Bacillus licheniformis. J Bacteriol. 1991 Aug;173(15):4725–4735. doi: 10.1128/jb.173.15.4725-4735.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lamb D. H., Bott K. F. Inhibition of Bacillus subtilis growth and sporulation by threonine. J Bacteriol. 1979 Jan;137(1):213–220. doi: 10.1128/jb.137.1.213-220.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LeDeaux J. R., Grossman A. D. Isolation and characterization of kinC, a gene that encodes a sensor kinase homologous to the sporulation sensor kinases KinA and KinB in Bacillus subtilis. J Bacteriol. 1995 Jan;177(1):166–175. doi: 10.1128/jb.177.1.166-175.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Martin I., Debarbouille M., Klier A., Rapoport G. Induction and metabolite regulation of levanase synthesis in Bacillus subtilis. J Bacteriol. 1989 Apr;171(4):1885–1892. doi: 10.1128/jb.171.4.1885-1892.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mizuno M., Masuda S., Takemaru K., Hosono S., Sato T., Takeuchi M., Kobayashi Y. Systematic sequencing of the 283 kb 210 degrees-232 degrees region of the Bacillus subtilis genome containing the skin element and many sporulation genes. Microbiology. 1996 Nov;142(Pt 11):3103–3111. doi: 10.1099/13500872-142-11-3103. [DOI] [PubMed] [Google Scholar]
  28. Mueller J. P., Bukusoglu G., Sonenshein A. L. Transcriptional regulation of Bacillus subtilis glucose starvation-inducible genes: control of gsiA by the ComP-ComA signal transduction system. J Bacteriol. 1992 Jul;174(13):4361–4373. doi: 10.1128/jb.174.13.4361-4373.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Noguchi N., Sasatsu M., Kono M. Genetic mapping in Bacillus subtilis 168 of the aadK gene which encodes aminoglycoside 6-adenylyltransferase. FEMS Microbiol Lett. 1993 Nov 15;114(1):47–52. doi: 10.1016/0378-1097(93)90140-w. [DOI] [PubMed] [Google Scholar]
  30. Ohnishi K., Hasegawa A., Matsubara K., Date T., Okada T., Kiritani K. Cloning and nucleotide sequence of the brnQ gene, the structural gene for a membrane-associated component of the LIV-II transport system for branched-chain amino acids in Salmonella typhimurium. Jpn J Genet. 1988 Aug;63(4):343–357. doi: 10.1266/jjg.63.343. [DOI] [PubMed] [Google Scholar]
  31. Parsek M. R., McFall S. M., Shinabarger D. L., Chakrabarty A. M. Interaction of two LysR-type regulatory proteins CatR and ClcR with heterologous promoters: functional and evolutionary implications. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12393–12397. doi: 10.1073/pnas.91.26.12393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schell M. A. Molecular biology of the LysR family of transcriptional regulators. Annu Rev Microbiol. 1993;47:597–626. doi: 10.1146/annurev.mi.47.100193.003121. [DOI] [PubMed] [Google Scholar]
  34. Slack F. J., Mueller J. P., Sonenshein A. L. Mutations that relieve nutritional repression of the Bacillus subtilis dipeptide permease operon. J Bacteriol. 1993 Aug;175(15):4605–4614. doi: 10.1128/jb.175.15.4605-4614.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Smith I., Paress P., Cabane K., Dubnau E. Genetics and physiology of the rel system of Bacillus subtilis. Mol Gen Genet. 1980;178(2):271–279. doi: 10.1007/BF00270472. [DOI] [PubMed] [Google Scholar]
  36. Stucky K., Hagting A., Klein J. R., Matern H., Henrich B., Konings W. N., Plapp R. Cloning and characterization of brnQ, a gene encoding a low-affinity, branched-chain amino acid carrier in Lactobacillus delbrückii subsp. lactis DSM7290. Mol Gen Genet. 1995 Dec 20;249(6):682–690. doi: 10.1007/BF00418038. [DOI] [PubMed] [Google Scholar]
  37. Sullivan M. A., Yasbin R. E., Young F. E. New shuttle vectors for Bacillus subtilis and Escherichia coli which allow rapid detection of inserted fragments. Gene. 1984 Jul-Aug;29(1-2):21–26. doi: 10.1016/0378-1119(84)90161-6. [DOI] [PubMed] [Google Scholar]
  38. Ward J. B., Jr, Zahler S. A. Regulation of leucine biosynthesis in Bacillus subtilis. J Bacteriol. 1973 Nov;116(2):727–735. doi: 10.1128/jb.116.2.727-735.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. van Rhijn P., Vanderleyden J. The Rhizobium-plant symbiosis. Microbiol Rev. 1995 Mar;59(1):124–142. doi: 10.1128/mr.59.1.124-142.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Lintig J., Kreusch D., Schröder J. Opine-regulated promoters and LysR-type regulators in the nopaline (noc) and octopine (occ) catabolic regions of Ti plasmids of Agrobacterium tumefaciens. J Bacteriol. 1994 Jan;176(2):495–503. doi: 10.1128/jb.176.2.495-503.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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